Anti-cancer Effects of Methotrexate in Combination With Melissa officinalis on HeLa Cancer Cell Line

Objectives: One of the well-accepted beliefs about natural products, considering the advances of recently appearing new edges and features of herbal medicine, is paying more attention to cancer treatments. However, they have not been properly studied with reasonable/reliable clinical trials in human subjects in most cases. Therefore, seeking in vitro effects of herbs like Melissa officinalis (MO) in cancer therapy to identify the involved possible mechanism in conjugation with configurative/morphological aspects of treated cells seems quite necessary. In this study, we evaluated the co-treatment effect of anti-cancer drug methotrexate (MTX) and MO on HeLa cancer cells. Methods: MTT assay was applied to assess the quantitative cytotoxicity effect of both MTX and Mo. Apoptosis assay via flow cytometry was used to determine the amount of apoptotic and necrotic cells. To further investigate the anti-cancer effects, DAPI staining and DNA ladder assays are used qualitatively to detect changes in the nuclei of cells that are a sign of apoptosis occurring and morphological modifications of DNA. Results: MTX and MO mixture showed high cytotoxicity and apoptosis rate compared to untreated cells. Furthermore, the morphological changes of MTX and MO mixture were more evident than that of single MO, MTX, and control groups. Conclusions: These data regarding cell growth reduction and apoptosis induction in HeLa cancer cells showed that MTX and MO mixture can be an appropriate platform for cancer therapy.